Regulation Of IGF-1 On Proliferation And Differentiation Of Sika Deer Antler Mesenchymal Cells

The antler horn is the only organ that can be fully regenerated in mammals.Therefore,antler is an ideal animal model for studying wound healing and mammalian organ regeneration,and it is a hot spot in regenerative medicine research.Mesenchymal cells have the characteristics of stem cells and can differentiate into cartilage tissue.During the growth and development of antler,antler mesenchymal cells can differentiate into chondrocytes and develop into chondrocytes under the induction of some factors of antler.In this experiment,sika deer was used as the research object.The effects of IGF-1 on the proliferation and differentiation of sika antler horn mesenchymal cells were studied by MTS,real-time PCR and flow cytometry.The regulation mechanism was also discussed.The results showed that IGF-1 significantly promoted the proliferation of antler mesenchymal cells,decreased the proportion of cells in G0/G1 phase,and increased the proportion of cells in S phase;while pretreatment with IGF-1 receptor inhibitor(PQ401)inhibited these effects..In order to further study the regulation of IGF-1 on the proliferation of antler mesenchymal cells,antler mesenchymal cells were isolated and cultured,and a certain concentration of IGF-1 recombinant protein and Notch blocker DAPT were added.The results showed that IGF-1 can significantly promote the proliferation of antler mesenchymal cells,reduce the proportion of G0/G1 phase,and increase the proportion of S phase;while pretreatment with Notch blocker DAPT can inhibit this effect.In order to study the regulation mechanism of IGF-1 on the differentiation of velvet antler mesenchymal cells,microsphere culture method was used to detect the concentration of IGF-1 recombinant protein and then detected by fluorescence quantitative PCR(Type II collagen,Col II).Proteoglycan(Aggrencan,AGC),cartilage oligomieric matrix protein(COMP)and SOX-9 expression changes.At the same time,the effect of IGF-1 on the expression of glycosaminoglycans in mesenchymal cells was detected by alexin blue staining.The results showed that the expression levels of Col II,AGC,COMP and SOX-9 were significantly increased in mesenchymal cells treated with IGF-1.After pretreatment with PQ401,IGF-1 inhibited Col II,AGC,COMP and The promotion of SOX-9 expression;IGF-1 treatment of mesenchymal cells increased glycosaminoglycan content,PQ401 pretreatment,inhibited this effect.Notch signaling can mediate the effect of IGF-1 on the differentiation of mesenchymal cells.After adding a certain concentration of IGF-1 recombinant protein and DAPT,the results showed that DAPT pretreatment inhibited IGF-1 against Col II,AGC,COMP and The effect of SOX-9 expression was also significantly reduced in mesenchymal cell glycosaminoglycan content.The effects of IGF-1 on superoxide anion and Gpx content in mesenchymal cells were further studied.The results showed that IGF-1 increased the content of superoxide anion and Gpx in mesenchymal cells,indicating that IGF1 may regulate cell oxidation.Effect to regulate the differentiation of mesenchymal cells.